Abstract:[Objective] The antimicrobial peptide, YFGAP from yellowfin tuna (Thunnus albacores), consisted of 32 amino acid residues, shows broad-spectrum antimicrobial activity against Gram-positive and Gram-negative bacteria without hemolytic activity. Clone and express the antimicrobial peptides, YFGAP in Escherichia coli BL21(DE3) and characterize its bioactivity. [Methods] Based on the amino acids of EK-YFGAP and L-EK-YFGAP, the genes were synthesized and ligated into pET-22b to construct these expression vectors, pET22b-ELP20-EK-YFGAP, pET22b-ELP40-EK-YFGAP and pET22b-ELP40-L-EK-YFGAP. The vectors were transformed into E. coli BL21(DE3) to express the fusion proteins. The fusion proteins were purified using the Elastin-like polypeptides (ELPs) as a purification tag by the method called inverse transition cycling (ITC) and digested by enterokinase. Then the recombinant YFGAP was purified using Vivaspin Turbo. Bioactivity of the peptide was tested. [Results] The fusion proteins, ELP40-EK-YFGAP and ELP40-L-EK-YFGAP were expressed and purified by two rounds of ITC. After digested by enterokinase, the recombinant YFGAP was purified using Vivaspin Turbo. Antimicrobial activity assay demonstrated the recombinant YFGAP exhibited high antibacterial activity against four fish pathogens without significant hemolytic activity. [Conclusion] The recombinant YFGAP was successfully expressed in E. coli using ELPs as a purification tag. This strategy does not require the use of chromatography, so that it is cost effective, easy to scale up and to multiplex. This study provides theoretical foundation and technical means for scale-up preparation of antimicrobial peptides by engineering method.

Abstract:[Objective] In recent decade, genome-scale metabolic network model (GSMM) has been flourishing rapidly. It has been an indispensable tool to investigate complex physiology of organisms through reconstruction of GSMMs and prediction cellular characteristics based on computational simulation. Visualization of simulation results can help to trace intuitively metabolic flux changes in the model and generate possible metabolic engineering strategies. [Methods] In this study, we first briefly summarized current methods for metabolic network visualization, followed by proposing a new method to realize visualization of GSMMs simulation based on Matlab. A pre-drawing of the metabolic network was performed first via CellDesigner software. Two functions of RAVEN toolbox were employed to integrate the raw map into Matlab and to realize visualization of simulation. [Results] As an example, the visualization of iYL619_PCP v1.7, a GSMM of Yarrowia lipolytica was realized. [Conclusion] According to the maps of visualization, we could monitor clearly flux changes in pathways under different environmental and genetic conditions to analyze efficiently simulation results.

Abstract:

Abstract:[Objective] To unveil effects of livestock grazing in Tibet on microbial interaction at the functional gene level. [Methods] We applied a recently developed network inference tool (Random Matrix Theory-based molecular ecological network) on GeoChip data related to carbon (C) and nitrogen (N) cycling with or without livestock grazing. [Results] C and N cycling gene networks in both control and grazing conditions had topological features of scale-free, small-world, modularity and hierarchy. Key genes in the grazing networks (hubs and connectors) differed substantially from those in the control. The grazing effects on soil microbial interactions were revealed by smaller, denser networks in the grazing samples, suggestive of environmental stress. In support of close linkages between aboveground plants and microbial community at this site, aboveground plant biomass was significantly (P=0.001) linked to grazing network topology. [Conclusion] Livestock grazing significantly altered microbial interaction at the functional gene level.

Abstract:

Abstract:[Objective] To solve the problem of contamination during ε-poly-L-lysine (ε-PL) fermentation, contamination microorganisms were screened and identified in this study. [Methods] Many strategies, like dilution coated plate, streak plate method, environmental stress method and liquid nutrient enrichment method, were performed to screen microorganisms from contaminated sample. Then the phylogenetic position of the isolate was determined through colony and microscopic morphology observation and 16S rRNA gene sequence analysis. Subsequently, the ε-PL tolerance of the isolate had evaluated. [Results] Contamination microorganisms were screened using liquid nutrient enrichment method isolated and identified as Acinetobacter bereziniae by 16S rRNA gene sequence analysis. Meanwhile, this isolate could tolerant high concentration of ε-PL. [Conclusion] Acinetobacter bereziniae was identified as the dominate contamination microorganism in ε-PL fermentation. And this find is helpful for the treatment of contamination microorganism in the process of ε-PL fermentation.

Abstract:[Objective] To understand the microbial community of denitrifying bacteria in Jiulong River estuary. [Methods] The sampling site was in Jiulong River estuary that was characterized by eutrophication. The water sample and sediment sample were collected and the environmental physicochemical factors were analyzed. The 16S rRNA gene and nirS gene clone libraries were constructed based on water sample and sediment sample respectively. [Results] In the 16S rRNA gene clone library, 86 valid sequences were grouped into 53 OTUs based on 97% sequence similarity and clustered into 6 different bacterial groups on the phylum level: Proteobacteria (Alpha, Beta, Gamma and Delta), Planctomycetes, Bacteroidetes, Actinobacteria, Firmicutes and Chloroflexi. Among these groups, Proteobacteria was the predominant member (62.9%). Of the nirS gene clone library 190 valid sequences were translated to conceptual amino acid sequences and group into 60 OTUs based on 82% similarity for genus identification. Among the 190 sequences, 71.6% were clustered into Proteobacteria phylum, including Alphaproteobacteria (5.8%), Betaproteobacteria (49.0%) and Gammaproteobacteria (16.9%). The most abundant OTU shared 100% sequence similarity to sequences from the nitrite reductase of Thauera sp. R-26906, a species of cultured denitrifier. [Conclusion] High diversity of the microbial community as well as nirS genes were found in the Jiulong River estuary. Most of the NirS sequences of this study shared quite high similarity to known sequences from various similar environments such as estuary and gulf.

Abstract:[Objective] Screening the typical, efficient desulfurization bacteria from the natural environment, exploring the growth characterization of the bacteria and conducting the preliminary desulfurization experiments. [Methods] The medium with sodium thiosulfate as the sole energy substrate was used to enrich desulfurization bacteria, and the pure strain was obtained after being streak plate culture and pure strain isolation for three times. Identifying the species of the strain by a variety of methods of Gram staining, plate colony morphologic observation, morphological characteristics research, 16S rRNA gene sequence analysis and the constructed molecular phylogenetic tree. [Results] Isolating a strain of efficient removal of thiosulphate bacteria, named CYJN-1 from the cooling water pool of Shanghai Waigaoqiao power station. The strain was Gram-negative bacteria, short rod and identified as Halothiobacillus neapolitanus (H. neapolitanus) bacteria. H. neapolitanus CYJN-1 had a strong adaptability to the changes of salinity, the salinity range for the growth of this bacteria was 0?5% (NaCl, W/V). Determine the optimum culture conditions strains: temperature 30 °C, pH 7.0, substrate concentration of 20 g/L. Under these conditions, the strain of sodium thiosulfate removal up to 98%. [Conclusion] Based on the high haloduric ability and the high ability of removal rate of the sodium thiosulfate, the CYJN-1 strain is of potential in the fields of biological desulfurization and biological metallurgy.

Abstract:[Objective] To isolate endophytic actinomycetes from Dongxiang wild rice (Oryza rufipogon), and to study the classification, antimicrobial activity and analysis the secondary metabolites from the high active strain S123. [Methods] S medium was used to isolate endophytic actinomycetes from Dongxiang wild rice. The isolates were classified by 16S rRNA gene sequence analysis. Antimicrobial activity of the isolates was tested using agar diffusion and growth rate method. Meanwile, to screen the potential activity compounds, The degenerated primer targeted on type-I polyketide synthase (PKS-I) was designed. Bioactivity strain S123 was subjected to large scale fermentation for isolating secondary metabolites. The compounds were separated and purified by various chromatographic methods, and the structure was determined by mass spectrum and nuclear magnetic resonance analyses. [Results] We obtained 11 actinomycete isolates from Dongxiang wild rice. The isolates belonged to 2 genera, Streptomyces (8) and Pseudonocardia (3). Among of these isolates 8 strains show their anti-microbial activities and 8 strains were positive for type-I PKS genes. Two compounds were isolated from strain S123 and determined as Nigericin and 17-O-demethylgeldanamycin separately. The compound Nigericin showed inhibition against Staphylococcus aureus, Bacillus subtilis and Rhizoctonia solani. [Conclusion] This is the first report of endophytic actinomycetes from Dongxiang wild rice and identified two activity compounds Nigericin and 17-O-demethylgeldanamycin associated with type-I PKS genes. It provides a reliable basis for studying Dongxiang wild rice endophytic actinomycetes diversity and seeking activity secondary metabolites.

Abstract:[Objective] A combination of Plackett-Burman Design (PBD) with Uniform Design (UD) was applied to optimize the nitrogen removal performance of immobilizing Micrococcus roseus. [Methods] Polyvinyl alcohol and Sodium alginate were used to immobilize cells. Using PBD, five factors including PVA, SA, temperature, the inoculation amount and the amount of pellet were selected from nine factors based on their significance (P<0.05). Next, UD was used to further optimize these five significant factors in order to improve nitrogen removal effect. Further, these data were analyzed by stepwise regression and second-order polynomial, which were based on Minitab and Matlab. [Results] The optimum conditions are as follows: PVA 9.6%, SA 2.2%, 33.4 °C, the inoculation amount 8%, the pellet number 500 per 100 mL, and the highest value of nitrogen removal rate peaked 60.9% after 72 h operation for nitrogen removal. [Conclusion] The combination of PBD with UD is an effective way to select significant factors for the optimization of denitrification performance of Micrococcus roseus. And Micrococcus roseus removes ammonia mainly through assimilation, nitrification and denitrification.

Abstract:[Objective] To better understand the behavior of microbial community during Chinese Maotai-flavor liquor fermentation, we studied the interaction between two functional strains, Saccharomyces cerevisiae and Bacillus licheniformis. [Methods] S. cerevisiae and B. licheniformis were tested in single and mixed culture. Biomass, ethanol and organic acid production were determined during the fermentation. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) coupled with matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) was used to analyze and identify differentially expressed protein of S. cerevisiae in single and mixed culture. [Results] S. cerevisiae showed a negative effect on B. licheniformis growth and the growth of itself was barely influenced. However, ethanol, pyruvic acid, lactic acid, malic acid, tartaric acid and succinic acid produced by S. cerevisiae in mixed culture have increased by 11.8％, 56.8%, 24.3%, 36.3%, 27.7% and 48.2% compared with its single culture, respectively. Citric acid yield was reduced by 35.1%. Proteomic profile of S. cerevisiae indicated that 69 spots (differential ratio>2) were differentially expressed in mixed culture. Among them, 24 protein spots were identified, including protein related to glycolysis, ethanol fermentation, cell wall integrity, and stress response. [Conclusion] B. licheniformis affected ethanol and organic acid metabolism of S. cerevisiae. That was significant for liquor quality and microbial interactions. Proteomic analysis of the responses of S. cerevisiae to mixed culture allows a deeper understanding of interaction mechanisms between both strains and will promote the understanding of the microbial community during Chinese Maotai-flavor liquor fermentation.

Abstract:[Objective] To explore the relationships between carotenoid (Car) structure and energy transfer efficiency in peripheral light-harvesting complex 2 (LH2) from anoxygenic phototrophic bacteria (APB). [Methods] Partially carotenoid-deficient LH2 (LC-LH2) was obtained from Rhodobacter azotoformans 134K20 by diphenylamine (DPA) inhibition. Three Cars including spheroidene (SE), rhodopin (RP) and okenone (OK) were obtained from three species of APB by thin layer chromatography and high performance liquid chromatography. The reconstitutions of different Cars into LC-LH2 were performed by using ultrasonic procedures in 10 mmol/L Tris-HCl buffer, pH 8.0, containing 0.1% LDAO. The structure and function of reconstituted LH2 (N-LH2) were studied by using UV-VIS spectra, Raman spectra and fluorescence spectroscopy. [Results] The deficiency of SE in LC-LH2 was about 64.7%. Three Cars with different conjugation length, substituent and polarity could be reconstituted into the partially SE-deficient LC-LH2, and the reconstitution efficiency was in the range from 24.0% to 29.4%, and the reconstitution efficiencies of SE and OK were higher than that of RP. Compared to the intrinsic planner conformation of SE in LC-LH2, the incorporated Cars also adopted a planner conformation in N-LH2. The Car to bacteriochlorophyll (BChl) energy transfer efficiency in different N-LH2 changed in the order of SE-LH2>RP-LH2>OK-LH2. The energy transfer efficiency in N-LH2 had a negative correlation with the conjugated length of Cars, whereas independent of the polarity of Cars. [Conclusion] In N-LH2, Cars bind to the apoproteins with a planner conformation, the conjugated length of Cars still plays a dominant role in controlling the Car to BChl energy transfer efficiency; however, the substituent and polarity of Cars have negligible effects on energy transfer efficiency.

Abstract:[Objective] Aim to study the effect of secretory aminopeptidase in Bacillus subtilis 168 on the growth, a gene knock-out mutant BS-AP-K was constructed. [Methods] Based on the Xer/dif recombinant system, the ywaD gene was deleted from the genome. The growth of wild type and BS-AP-K in different media was monitored. [Results] It turned out that the deletion of secretory aminopeptidase had a negative effect on the growth of the strain. The impaired growth rate of BS-AP-K strain can be overcome by amino acid supplements. [Conclusion] This study indicated that the secretory aminopeptidase played an important role in supplying amino acids required for growth by hydrolyzing proteins or peptides.

Abstract:[Objective] A luminous Basidiomycete, designated MRC-lf3, was isolated from Fujian, China, and its taxonomic status was confirmed, and its life cycle was recorded. [Methods] The mycelial pure culture was obtained by tissue isolation method; morphological characteristics and the phylogenetic analysis of ITS rDNA sequence were used to analyze the taxonomic status of this fungus; the morphological and luminous characteristics were recorded by Digital Single Lens Reflex. [Results] Both morphological characteristics and ITS rDNA phylogenetic analysis results demonstrated that the strain MRC-lf3 was Neonothopanus nambi. Our study also finding that both ability to emit light and color of the fruiting body could be changed by different illumination intensity. [Conclusion] We isolated the luminous fungal strain from Fujian China, which was identified as Neonothopanus nambi, this is the ?rst record of genus Neonothopanus in China. This new record of Neonothopanus nambi brings the total number of luminous fungi in the Chinese Mainland to 8 species.

Abstract:[Objective] Identification specimen Dxhir140901 which parasitized on Lepidopterous larvae. [Methods] The comparison of morphological characteristics and the analysis of phylogenetic tree and the evolution network based on internal transcribed spacer (ITS1-5.8S-ITS2rDNA) sequence were used for identification of this specimen. [Results] Morphological observation: morphological characteristics of the strains Gzuifr-hir140901 was similar to Hirsutella Pat. It has two types of phialides, A-phialides, column, (1.8?6.3) μm×1.8 μm. B-phialides, columnar base, awl-shape and tapers, width of base 3?3.8 μm, length 21?63 μm, width of neck 1.8?2.0 μm, Phialides formed directly from the end of mycelium. Conidia orange segments or oval, (8.1?10.8) μm×(2.7?5.4) μm, mucus thick 1.8?2.7 μm. From the results of the phylogenetic analysis we can find that the Gzuifr-hir140901 strains and Ophiocordyceps macroacicularis S. Ban, T. Sakane & Nakagiriin belong to the same clade with a 98% approval rating. Branching sequence analysis supports the same results. [Conclusion] The sample Dxhir140901 and strains Gzuifr-hir140901 was a anamorph of O. macroacicularis which is a new record from China.

Abstract:[Objective] Gene knockout mutant Δ0448 and the functional complementary strain CΔ0448 were constructed to investigate the role of SSU0448 gene on Streptococcus suis serotype 2 (SS2) morphology and virulence. [Methods] Based on the principle of homologous recombination, the SSU0448 gene was knocked out in SS2 virulent strain 05ZYH33. PCR analysis was carried out to identify the mutant, then screened the functional complementary strain CΔ0448 containing the SSU0448 gene expressing plasmid. The morphology and virulence of the established strains were examined. [Results] PCR analysis and DNA sequencing identified the knockout mutant Δ0448 and the SSU0448 gene was completely replaced by the spcr cassette. The functional complementary strain CΔ0448 was also successfully constructed. Gram staining showed that the mutant Δ0448 grew much shorter than the wild type 05ZYH33 and faster to reach the plateau phase. Deletion of the SSU0448 gene did not change the virulence of SS2 in mice infection assays. [Conclusion] Knockout of SSU0448 gene obviously influenced the chain ability of Streptococcus suis serotype 2, and its deficiency has no significant effect on the virulence in the mice model while it may delay the pathogenesis.

Abstract:[Objective] Objectives of this study were to further understand the mechanism on phosphate solubilization of Rahnella sp. W25 and its solubilization ability under buffering condition. [Methods] Regulating pH of nutrient solution was conducted in the process of liquid shake flask culture to research the influence of buffer capacity on the phosphate solubilizing ability of Rahnella sp. W25 under simulation soil condition. Research the solubilizing ability and acetic-producing characteristics under different carbon and phosphorus sources condition though single factor text and HPLC. [Results] The result showed that the available phosphorus content reached up to its maximum after cultivated 120 h in the solution of tricalcium phosphate. The change of available phosphorus content and pH in solution showed significant negative correlation (P<0.01). At cultivated 48?96 h, strain W25 had stronger buffering capacity and the difference of available phosphorus content between regulating pH using and without using NaOH was not significant (P<0.05). Strain W25 weaken after cultivated 120 h and basically lost buffering capacity after cultivated 168 h. The solubilizing ability showed significant different under different carbon sources (P<0.05). The order of solubilization ability as follows: glucose>lactose>sucrose>mannitol>starch. It also showed extremely significant different under different phosphorus sources (P<0.01). The order was tricalcium phosphate>iron phosphate>aluminum phosphate>ground phosphate rock. The type and concentration of organic acids were dissimilar under different carbon and phosphorus sources conditions. The solubilization ability of strain W25 was both related to the type and concentration of organic acids. [Conclusion] Results of this study could provide the condition of deeply researching mechanism on phosphate solubilization and the theoretical basis of application of Rahnella sp..

Abstract:[Objective] We established the mutation library of pathogenic Vibrio parahaemolyticus (Vp), analyzed the phenotype characteristic of hemolytic ability changed mutations, to provide the research basis for deeper exploration and understanding of the regulation system and mechanism of tdh. [Methods] In this study, the Mini-Tn5-Km2 was inserted randomly into the genome of pathogenic Vp (ATCC 33846) by biparental mating. The conjugants (KmRAmpS) were investigated using thiosulfate citrate bile salts sucrose agar medium with kanamycin (Km) and ampicillin (Amp) respectively. After detection of the Km gene by polymerase chain reaction, library of mutations was established, which the Tn5 was inserted in different gene locus randomly. The hemolytic activity was evaluated with human blood agar. Different characters of these conjugants were evaluated, including growth rate, biofilm formation and motility ability. [Results] Library of mutations was successfully established including 490 mutants. After evaluating hemolytic activity, the phenotype of 5 strains was changed, including 2 up-regulated strains and 3 down-regulated strains. There was significant difference between the 5 conjugants and parent strain on growth rate, biofilm formation and motility ability. Among 3 down-regulated strains, the biofilm formation ability of 2 conjugants was increased (P<0.05). The residual 3 strains demonstrated the significant difference (P<0.05) on growth rate, biofilm formation ability and motility, comparing with their parent strain. [Conclusion] Tn5 transposon can be used to establish the library of Vp mutations; Hemolytic ability of Vp is related to the phenotype; The five conjugants will be contributed on investigation of regulation system and mechanism of tdh in further study.

Abstract:[Objective] The aim of this study was to screen strains with high penicillin G acylase (PGA) activity. [Methods] Induced mutation of Bacillus megaterium ATCC 14945 was performed by LiCl-ultraviolet composite mutagenesis and atmospheric and room temperature plasma (ARTP) mutagenesis. Strains treated by two mutation methods were spread on solid plate culture medium. The colony growing on plate was inoculated into liquid medium and cultured at 28 °C, 250 r/min. Continuing culture was performed via transferring bacterium broth to fresh liquid medium with 10% of inoculum size to obtain second generation broth. After 6 h, phenylacetic acid was added with 0.1% final concentration into second generation broth and cultured for 40 h. The bacterium broth was centrifuged for 5 minutes with 8 000 r/min. The supernatants are crude enzyme and their PGA activities were assayed with NIPAB method. The enzyme-producing conditions, including the addition quantity and timing of phenylacetic acid were optimized using the strain with the best PGA enzyme of activity from the mutants. The protein properties of PGA from the strains induced before and after was analyzed by SDS-PAGE. [Results] The strains numbered 12-4 had the highest PGA of 39.60 U/mL. PGA activity increases by 8.5 times than that of original strains. After culturing strains12-4 for 6 h and adding phenylacetic acid into broth with quantity of 0.2% (W/V), continuing culture for 50 h, the PGA activity from the supernatants reached to 78.45 U/mL, which is 16.8 times than that of original strains. PGA from the strains induced before and after all has α and β subunit. The amount of α subunit in PGA from the induced strains was no significant change. However, the amount of β subunit significantly increased, also protein bands locating between α and β subunit markedly increased. [Conclusion] The activity of PGA from Bacillus megaterium could be increased by mutation. The obtained strains and PGA-producing conditions in this study are of important value to industrial production.

Abstract:[Objective] To investigate the role of kdpD gene in Vibrio alginolyticus, a kdpD gene deletion mutant was generated. [Methods] Combining with chloramphenicol (Cm) and sucrose screening, we constructed an in-frame deletion mutant of kdpD using overlap PCR and homologous recombination technology. We then investigated the effect of the mutant on its growth, ECPase activity and virulence in fish. [Results] The mutant ΔkdpD was successfully constructed. The mutant ΔkdpD showed no differences of ECPase activity and growth from the wild-type strain. But a decrease in swarming ability and biofilm formation was found. Moreover, virulence assays with zebrafish model confirmed that virulence of the mutant declined by 8.84 times. [Conclusion] The kdpD gene could regulate swarming ability, biofilm formation and virulence, but it has no effect on growth and ECPase activity.

Abstract:[Objective] The purpose of this study was to investigate the distribution, diversity and antimicrobial activity of endophytic bacteria of Lycium barbarum L. of Ningxia. [Methods] The 16S rRNA gene sequencing and agar diffusion were employed to explore the diversity and antimicrobial activity of endophytic bacteria isolated from different organs and tissues of Lycium barbarum L.. [Results] Total of thirty-four isolates were obtained. The result of diversity analysis showed that these isolates could be phylogenetically classified into 11 genera and 7 families, based on their 16S rRNA gene sequencing. The quantity and clonal composition of bacterial communities were distinct from various tissues. The quantity of endophytic bacteria in the different tissues was in order of roots > leaves > flowers > fruits; the diversity of bacteria was flowers > roots > leaves > fruits. Additionally, Bacillus spp. was a predominant species and was isolated in all tested tissues. Analysis of antimicrobial activity demonstrated that 76.5% of isolates had an antimicrobial activity against at least one of the five tested pathogenic bacteria, among which Bacillus R2, R7, L3 and Brevundimonas R3 displayed significant antimicrobial activity against the plant pathogens Colletotrichum nigrum and Exserohilum turcica. Of note, the majority of isolates showed weak inhibition effect against Escherichia coli and Staphylococcus aureus. [Conclusion] Endophytic bacteria of Lycium barbarum L. are genetically diverse and most of them have a strong inhibitory activity against plant pathogenic fungi.

Abstract:[Objective] To analysis the molecular genetic characteristics of influenza A virus H3N2 and N7N9 viruses in one case of mixed infection. [Methods] Using a set of primers and probes for influenza virus genotyping, specimens were detected by real-time quantitative PCR method. Whole-genome sequences of virus isolates were obtained by second-generation sequencing technology. [Results] In April 2013, one mixed infection with seasonal H3N2 influenza virus and avian influenza H7N9 virus were confirmed in Nanjing, and two mixed viruses were isolated, which named A/Nanjing/M1/2013 (H3N2) (M1-H3N2) and A/Nanjing/M2/2013 (H7N9) (M2-H7N9), respectively. Some important molecular markers associated with host adaptation and virulence were identified in M2-H7N9 virus. M2-H7N9 virus had substitution Q226L (H3 numbering) in the receptor-binding site of hemagglutinin, which may facilitate the viruses to bind to sialic acid (SA)-2,6-Gal-terminated saccharides that are abundant in human upper respiratory epithelium. There was substitution at position 627 (E→K) in the PB2 protein, which enhances the ability of the virus infection on the human. [Conclusion] This study provided a direct evidence for human as “mix vessel” of influenza virus, and the reassortment of seasonal influenza viruses and avian H7N9 viruses should be concerned.

Abstract:Oncolytic virus (OV) is a class of antitumor agents that selectively infect and kill tumor cells while sparing normal cells. Among various oncolytic viruses, oncolytic herpes simplex virus type 1 (OHSV-1) is one of the most widely explored agents. OHSV-1 can be constructed through various strategies. It has been investigated in clinical trials for patients with many types of cancers. These clinical studies have shown the safety and efficacy of OHSV-1. This review briefly summarizes the molecular biologic characteristics and advantages of OHSV-1, the development and targeting strategies of OHSV-1, the progresses in research on various OHSV-1 and discusses the currently problems with OHSV-1 in cancer therapy.

Abstract:Based on the author’s teaching practice of “Microbiology” in Shanghai Jiao Tong University, this paper discuss and explore the application of a “Knowledge Organization” teaching strategy. The knowledge organization teaching strategy can help teachers to eliminate “blind spots” before class, guide students to form their own knowledge organization and knowledge structure, and improve students’ comprehensive ability to solve complex microbiological problems.

Abstract:The modular teaching pattern for microbiology experiments helps to improve undergraduates’ experimental skills and their comprehensive application ability. The authors described the construction of the modular microbiology experiment course and its application. First, we established a comprehensive, research-based modular teaching system. The system is composed of three modules including investigation of the environmental microbial diversity; screening and isolation of the special functional microbes, culture conditions optimization and strain improvement; the microbiological testing of water. There were also a total of 12 experimental units. The contents are designed to: (1) Emphasize the training of students in their basic microbiology experimental skills and guide their learning; (2) Strengthen the connection between experiment units and the coherence of different modular experiments; (3) Refine the experimental contents, simplify the experimental steps and reduce students’ study load. Second, we organized the seminars to summarize and gain the experience of teaching, discuss and optimize the following unit plans. These seminars will help us to improve the teaching effect and the teacher’s guidance ability in a microbial experiment course. Third, we established different ways to evaluate the students’ learning, such as routinely reviewing their work for each experimental unit, implementing a practical examination to test their experimental skills at the end of the semester, reviewing their ability to write reports for experimental units, as well as writing for modules in a research article format. All together these will result in better evaluations on the students' comprehensive ability. As an incentive, we will also exhibit the outstanding achievements produced in each of the experimental courses.